論文 - 詳細
| RRC ID | 85158 |
|---|---|
| 著者 | Schreiber T, Prange A, Schäfer P, Iwen T, Grützner R, Marillonnet S, Lepage A, Javelle M, Paul W, Tissier A. |
| タイトル | Efficient scar-free knock-ins of several kilobases in plants by engineered CRISPR-Cas endonucleases. |
| ジャーナル | Mol Plant |
| Abstract |
In plants and mammals, non-homologous end-joining is the dominant pathway to repair DNA double-strand breaks, making it challenging to generate knock-in events. In this study, we identified two groups of exonucleases from the herpes virus and the bacteriophage T7 families that conferred an up to 38-fold increase in homology-directed repair frequencies when fused to Cas9/Cas12a in a tobacco mosaic virus-based transient assay in Nicotiana benthamiana. We achieved precise and scar-free insertion of several kilobases of DNA both in transient and stable transformation systems. In Arabidopsis thaliana, fusion of Cas9 to a herpes virus family exonuclease led to 10-fold higher frequencies of knock-ins in the first generation of transformants. In addition, we demonstrated stable and heritable knock-ins in wheat in 1% of the primary transformants. Taken together, our results open perspectives for the routine production of heritable knock-in and gene replacement events in plants. |
| 巻・号 | 17(5) |
| ページ | 824-837 |
| 公開日 | 2024-5-6 |
| DOI | 10.1016/j.molp.2024.03.013 |
| PII | S1674-2052(24)00086-8 |
| PMID | 38520090 |
| MeSH | Arabidopsis / enzymology Arabidopsis / genetics CRISPR-Cas Systems* / genetics Endonucleases / genetics Endonucleases / metabolism Gene Knock-In Techniques* Nicotiana* / genetics Plants, Genetically Modified Triticum / genetics |
| IF | 12.084 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 301 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| コムギ | LPGKU2304 |